This study was carried out for the classification of mycobacteria using the thin-layer chromatography of mycobacterial lipids. Results as follows: Of the 12 strains of mycobacteria, the two spots on chromatogarphy were three strains of mycobacteria(BCG, M. tuberculosis($H_{37}J$) and M. ulcerans) and three spots on chromatography were two strains of mycobacteria(M. kansasii and M. balnei). The method of thin-layer chromatography of mycobacterial lipids was considered to capable for the use of classification of mycobacteria.